Confirmatory cultural and gene-detection studies: an examination

'La théorie des germes de Louis Pasteur est une fiction ridicule.' Pierre Pachet, professor of physiology, Toulouse (1872)

The evidence for a causal association between a potentially treatable Chlamydia pneumoniae infection and majority subsets of Multiple Sclerosis, as detailed on the first page, is very strong. It is far stronger than hypothesis. Why, then, is this evidence almost unknown within the medical establishment? MS is often a devastating illness which usually strikes during the most productive years. It is the commonest neurological illness in young adults. No present treatment is anywhere near satisfactory, and, as the disease becomes progressive, present-day medicine has little to offer.

The problem is one of collation. A multitude of scientific studies - some of great elegance - can be linked together to support the causal association very powerfully, but this information has not been brought together in a broad, comprehensive manner within a widely-read multidisciplinary medical journal. The reason for this delay is the process by which scientific papers are selected for publication.

Editors must be selective. Scientific studies vary in their quality. No editor would wish to publish worthless material. At the same time no editor can be expected to possess the expertise necessary to evaluate the material he or she receives. One of the editorial staff will read the manuscript. If it is obviously poor then it will be filed in the basement and a polite note of rejection will be sent to the author. If it passes preliminary scrutiny as being reasonable and worthwhile, copies will be sent to two or three referees who are considered knowledgeable in the subject. The referees will read the manuscript and submit a report. This is the peer-review system, and is generally considered to be the least bad way of selecting scientific work for publication. Its advantages are that it can pick up sloppy or improbable work; work which places hypotheses upon hypotheses, or work which does not stand up to analysis.

It has drawbacks, though. The biggest drawback is that it resists any change to the underlying Weltanschauung [1] or 'mindset' on which the subject rests. The peers are senior establishment figures; their reputations are built solidly on a thesis of ideas which they share with each other. Thus it is that any work which casts doubt on the validity of the prevailing ethos is likely to be treated very harshly. Its principles are likely to be pulled to pieces in an unfair way; its findings, even when clear, are made to sound doubtful, and the argumentation in the discussion made to sound improbable. None of this is (one hopes) conscious. (A few days ago I happened to bring up this subject on meeting a friend in the hospital; she immediately took a copy of the Guardian from under her arm and said, 'you might like to have a look at this.' It was an article by Cordelia Fine. It is indeed apposite. I've put a few quotes at the bottom of the page.)

In the case in point the 'peers' are dedicated to the theory of inflammatory autoimmunity as the primary flywheel of the disease. They have made an enormous investment into this: years of their lives, ongoing research projects, academic reputations, lucrative connections to pharmaceutical companies eager to profit by expensive agents such as monoclonal antibodies. No-one is going to fund a large trial of out-of-patent antibiotics and unpatentable supplements. The establishment wishes that any idea of infection being at the root of MS would go away. That is why, if you study the literature, you'll find that negative studies [2] are given equal weight with studies that detected Chl pneumoniae in the CSF, and grew the organism and found specific antibodies [3]. Another (again one hopes) unconscious method of down-rating evidence which counters the established view is to side-line those who have made the positive discoveries. 'I know that Chlamydia pneumoniae has been found in the CSF of people with MS,' said a world-respected MS neurologist recently, 'but those early findings have never been repeated.' He is stating a half-truth, but only a half-truth.

Those early findings have indeed been repeated. There are, in fact, many good, worldwide, confirmatory studies which have detected Chl pneumoniae in the CSF of persons with with MS by culture or specific gene detection. Here is a précis.

The preliminary work was carried out by workers at Vanderbilt University, Nashville, TN in the US. 27 patients with MS and 27 patients with other neurological disease were studied. Polymerase chain reaction assays demonstrated the presence of C pneumoniae MOMP gene in the CSF of 97% of MS patients versus 18% of patients with other neurological disease (OND) [3] The same workers isolated C pneumoniae from the CSF in 64% of MS patients and 11% of OND controls. 86% of MS patients had increased CSF antibodies to C pneumoniae elementary body antigens as shown by enzyme-linked immunosorbent assay absorbance values that were 3 SD greater than those seen in OND controls. The specificity of this antibody response was confirmed by western blot assays of the CSF, using elementary body antigens. Moreover, CSF isoelectric focusing followed by western blot assays revealed cationic antibodies against C pneumoniae. The authors are careful to say that this gives evidence for but does not prove and aetiological link.

In 2000 Layh-Schmitt and colleagues in Heidelberg, Germany, reported the presence of Chl pneumoniae in 5 of 10 patients with MS using PCR. In a further study they detected the organism in 2 of 20 patients with MS, 3 of 17 patients with possible/probable MS and 0 of 56 patients with other neurological disease (OND) [4].

In 2001 Ikejima and colleagues, in the USA, reported the presence of Chl pneumoniae genes in the CSF of 11 of 16 patients with MS. These workers used a commercial kit rather than an in-house method [5].

Also in 2001 Geiffers and colleagues in Lubeck, Germany, detected Chl pneumoniae DNA in the CSF of 12 of 58 patients with MS and 20 of 47 patients with OND. However, Chl pneumoniae DNA was not detected in the CSF of any of 67 neurologically healthy persons. These authors conclude that Chl pneumoniae is probably a highly neurotropic pathogen [6.]

In 2002 Hao and colleagues, in Japan, reported the presence of Chl pneumoniae in 9 of 28 patients with MS and in 2 patients with inflammatory disease in 15 patients with OND. Elevated titres of specific antibody was found in the CSF of 42 of 84 patients with MS and in 28% of patients with OND [7.] [] Interestingly these workers observed a higher rate of Chl pneumoniae gene transcription in the patients with MS; this suggests a more active bacterial metabolism, and may point to host factors playing a part in the disease [7.]

In 2003 Grimaldi and colleagues, in Italy, detected Chl pneumoniae gene-sequences in the CSF of 23 of 107 patients with MS and 2 of 77 patients with OND [8].

In 2003 another study from the Heidelberg group detected the chlamydia-specific ompA gene in the CSF of 50% of patients with MS and in 28.1% of 89 OND patients [9.] In a most interesting further study, these authors found chlamydial HSP60-mRNA in the CSF of 75% of patients with MS, indicating active bacterial activity: chlamydiae are known to upregulate HSP60. This has the effect of preventing host cell apoptosis.

In 2002 a conjoint study of Chlamydia pneumoniae gene-sequences in the CSF was undertaken by workers at Vanderbilt University Medical Centre and the University of South Florida. The two laboratories used different DNA extraction and PCR techniques. In definite MS patients the detection rates were 72% (VUMC) and 61% (USF); in probable monosymptomatic patients the detection rates were 41 and 54% respectively; in OND controls the signal was positive in 7% and 16% respectively [10].

This repeated confirmation by different centres - using different methodology and targets - leaves little room for doubt about the association of Chl pneumoniae with MS. We have recently reviewed the evidence in some detail [11.]

Those who have read the above compilation will note, in the PCR findings within the CSF of persons with MS, a wide range of results, ranging from 10% to 97% positive signal. What is the cause of this diversity? Three possibilities might be considered: differences in methodology; geographical differences in strains; very small organism numbers. A very clear account of PCR in the detection of bacterial gene-sequences in the CSF is given by Yamamoto, who brings clarity to a difficult subject [12.] One of the most problematic issues, this paper says, is that of organism numbers. For one thing, Chl pneumoniae, under chronic stress conditions, drastically curtails EB production. Extracellular EB nucleic acids and proteins are thus likely to be present in minute quantities within the CSF, and, furthermore, are likely to be swept away very rapidly. The CSF is often considered to be a slow-moving - even sluggish - stream; this is not the case. The body of fluid is exchanged some three times in a 24 hour period; the volume of the CSF-containing cavities is about 150ml and approximately 500ml of CSF is produced every day. The anatomy of the CSF circulatory pathway is such that there are no 'dead spaces'. EBs will be quickly removed. The determination of positivity or negativity may well lie at the edge of chance; any slight advantage of methodology would be critical. It is possible that EB release may be intermittent. It will be noted that the positive reports are from centres round the world; little is known about geographical variation in the external proteins of the EB, or whether there are functional strain-differences; some strains might, for instance, produce more EBs than others while in the persistent state. An elucidation of the possible events which may happen in the course of persistent infective states with this organism will present one of the greatest challenges in medical microbiology in this century.

The information is present, but is not as yet collated. This is the reason why, if you want to stand a chance of recovery from MS, you will have to trawl through the millions of pages of journals listed in PubMed, using appropriate search terms: and when you find the material you will have to edit it to remove the establishment bias. Or else find someone who has done this for you and who has put the information on the Internet.

Speaking of the Internet, it is worthwhile examining the bona fides of any web-forum in which you may wish to participate. Some are sponsored by business organisations which do not permit discussion antithetical to their commercial interests. Posts which allude to treatable infection as a component of the MS syndrome tend to be removed. and can be recommended as being forums which allow truly independent discussion.

Notes and References

1. Weltanschauung: the accepted scheme of things or an 'axiomatic framework'. It is usually translated as 'world view' and refers to a person's or group's culturally constructed version of reality. A term given philosophical currency by the phenomenologist Edmund Husserl.

2. Boman J, Roblin PM, Sundstrom P, Sandstrom M, Hammerschlag MR. Failure to detect Chlamydia pneumoniae in the central nervous system of patients with MS. Neurology. 2000 Jan 11;54(1):265.

3. Sriram S, Stratton CW, Yao S, Tharp A, Ding L, Bannan JD, Mitchell WM. Chlamydia pneumoniae infection of the central nervous system in multiple sclerosis. Ann Neurol. 1999 Jul;46(1):6-14.

4. Layh-Schmitt G, Bendl C, Hildt U, Dong-Si T, Juttler E, Schnitzler P, Grond-Ginsbach C, Grau AJ. Evidence for infection with Chlamydia pneumoniae in a subgroup of patients with multiple sclerosis. Ann Neurol. 2000 May;47(5):652-5.

5. Ikejima H, Haranaga S, Takemura H, Kamo T, Takahashi Y, Friedman H, Yamamoto Y. PCR-based method for isolation and detection of Chlamydia pneumoniae DNA in cerebrospinal fluids. Clin Diagn Lab Immunol. 2001 May;8(3):499-502.

6. Gieffers J, Pohl D, Treib J, Dittmann R, Stephan C, Klotz K, Hanefeld F, Solbach W, Haass A, Maass M. Presence of Chlamydia pneumoniae DNA in the cerebral spinal fluid is a common phenomenon in a variety of neurological diseases and not restricted to multiple sclerosis. Ann Neurol. 2001 May; 49(5):585-9.

7. Hao Q, Miyashita N, Matsui M, Wang HY, Matsushima T, Saida T. Chlamydia pneumoniae infection associated with enhanced MRI spinal lesions in multiple sclerosis. Mult Scler. 2002 Oct;8(5):436-40.

8. Grimaldi LM, Pincherle A, Martinelli-Boneschi F, Filippi M, Patti F, Reggio A, Franciotta D, Allegra L, Comi G, Blasi F. An MRI study of Chlamydia pneumoniae infection in Italian multiple sclerosis patients. Mult Scler. 2003 Oct;9(5):467-71.

9. Dong-Si T, Weber J, Liu YB, Buhmann C, Bauer H, Bendl C, Schnitzler P, Grond-Ginsbach C, Grau AJ. Increased prevalence of and gene transcription by Chlamydia pneumoniae in cerebrospinal fluid of patients with relapsing-remitting multiple sclerosis. J Neurol. 2004 May; 251(5): 542-7.

10. Sriram S, Yao SY, Stratton C, Calabresi P, Mitchell W, Ikejima H, Yamamoto Y. Comparative study of the presence of Chlamydia pneumoniae in cerebrospinal fluid of Patients with clinically definite and monosymptomatic multiple sclerosis. Clin Diagn Lab Immunol. 2002 Nov; 9(6): 1332-7.

11. Stratton CW, Wheldon DB. Multiple sclerosis: an infectious syndrome involving Chlamydophila pneumoniae. Trends Microbiol. 2006 Nov;14(11):474-9.

12. Yamamoto Y. PCR in diagnosis of infection: detection of bacteria in cerebrospinal fluids. Clin Diagn Lab Immunol. 2002 May;9(3):508-14.

'So evidence that fits with our beliefs is quickly waved through the mental border control, while counter-evidence must submit to close interrogation and, even then, will probably not be admitted. As a result, people can end up holding their beliefs even more strongly after seeing the counter-evidence. . .

'Part of this attachment may be because there is a sense in which our important beliefs are an integral part of who we are. To bid a belief adieu is to lose a cherished part of our identity. . .

[Cordelia Fine: The Vain Brain The Guardian 26th January 2006 Pp G 2 16 - 17 ]


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page created 28th January 2006; last edited 18th March 2011